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Kit for detecting content of MR-1S (Myofibrillogenesis Regulator-1S) in human plasma and preparation method thereof

A MR-1S and human detection technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as no public reports, achieve high sensitivity, good clinical application prospects, and meet the needs of clinical diagnosis and treatment monitoring.

Inactive Publication Date: 2012-03-07
上海永昶医学诊断用品有限公司 +1
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

But so far, MR-1S, as a potential tumor marker with good clinical application prospects, has not been publicly reported so far.

Method used

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Experimental program
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Effect test

preparation example Construction

[0049] Preparation of human MR-1S fusion protein: extract the blood genomic DNA of ovarian cancer patients, and use the method of molecular cloning, that is, amplify the target gene human MR-1S from the blood genomic DNA by PCR, and digest it with the same double enzyme Cut prokaryotic expression vector pET21a (+) or PGEX-4T-1, construct pET21a-MR-1S or PGEX-4T-1-MR-1S recombinant plasmid, transform Escherichia coli BL21, induce with isopropylthiogalactoside IPTG MR-1S-His fusion protein or MR-1S-GST fusion protein expression, Western blot identification after affinity purification, MR-1S-His fusion protein or MR-1S-GST fusion protein concentration, with 10mmol / L, pH7. 2 ℃ of PBS was fully dialyzed and activated, freeze-dried into freeze-dried powder to obtain human MR-1S fusion protein, and stored at -20°C.

[0050] Preparation and purification of anti-human MR-1S antibody: Immunize experimental animals with the human MR-1S fusion protein purified above to prepare specific hi...

Embodiment 1

[0052] 1. First prepare the following reagents:

[0053] 1. One anti-human MR-1S antibody pre-coated plate, the coating in the anti-human MR-1S antibody pre-coated plate is anti-human MR-1S antibody with a concentration of 50-500 μg / ml;

[0054] 2. One bottle of anti-human MR-1S antibody enzyme labeling solution;

[0055] 3. Human MR-1S standard solution with serial concentrations;

[0056] 4. The positive quality control solution is phosphate solution PBS containing human MR-1S fusion protein, and the negative quality control solution is 0.9% physiological saline. The concentration of the human MR-1S fusion protein is 300 ng / ml; the concentration of the phosphate solution PBS is 0.01mol / L, and the pH value is 7.4;

[0057] 5. Washing solution: Tris 2.42 g, 0.1 mol / L HCl 13 ml, Tween-20 0.5 ml, add distilled water to 1000 ml;

[0058] 6. Sample diluent: KH 2 PO 4 0.2 g, Na 2 HPO 4 12H 2 O 2.9 g, NaCl 8.0 g, KCl 0.2 g, Tween-20 0.5 ml, bovine serum albumin BSA 10 g, a...

Embodiment 2

[0100] 1. First prepare the following reagents:

[0101] 1. 1 piece of MR-1S antigen pre-coated plate, the coating material in the MR-1S antigen pre-coated plate is 1~5 μg / ml human MR-1S fusion protein as antigen;

[0102] 2. One bottle of anti-human MR-1S antibody enzyme labeling solution;

[0103] 3. Human MR-1S standard solution with serial concentrations;

[0104] 4. Positive quality control solution: phosphate solution PBS containing human MR-1S fusion protein; negative quality control solution: 0.9% physiological saline, the concentration of the human MR-1S fusion protein is 300 ng / ml; The concentration of described phosphate solution PBS is 0.01mol / L, pH value is 7.4;

[0105] 5. Washing solution: Tris 2.42 g, 0.1 mol / L HCl 13 ml, Tween-20 0.5 ml, add distilled water to 1000 ml;

[0106] 6. Sample diluent: KH 2 PO 4 0.2 g, Na 2 HPO 4 12H 2 O 2.9 g, NaCl 8.0 g, KCl 0.2 g, Tween-20 0.5 ml, bovine serum albumin BSA 10 g, add distilled water to 1000 ml;

[0107] 7...

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Abstract

The invention belongs to the technical field of biomedical inspection, and particularly relates to an enzyme-linked immunosorbent assay kit for detecting the content of Myofibrillogenesis Regulator-1S (MR-1S). The enzyme-linked immunosorbent assay kit consists of the following reagents: an anti-human MR-1S antibody-precoated microplate or MR-1S antigen-precoated microplate; a bottle of anti-human MR-1S antibody enzyme-marked solution; MR-1S standard solutions of serial concentrations; a bottle of positive quality control solution; a bottle of negative quality control solution; a bottle of normal saline; cleaning mixture; sample diluent; blocking solution; substrate solution; and stop solution. Compared with the prior art, the invention has the advantages that: the detected result of the established kit shows that the kit can quantitatively detect the concentration of MR-1S, and the detected result can reflect the change range and the development trend of human MR-1S and meets the requirements of clinical diagnosis and treatment monitoring, moreover, an assay can be fulfilled in 120 minutes, the reagents and the method are stable, and are easy to operate, the sensitivity is high, the specificity is stronger, and the kit has a better clinical application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedical testing, and in particular relates to an enzyme-linked immunosorbent assay kit for detecting the content of S subtype human myofibrogenesis regulatory factor 1 and a preparation method thereof. Background technique [0002] Currently, the morbidity and mortality of malignant tumors are increasing year by year. At present, tumor diagnosis mainly relies on imaging examination, cytology, biochemistry and immunology tests. The latter mainly examines tumor markers (tumor markers) in serum or plasma, referred to as TM. Because the detection of TM is more economical, convenient to obtain materials, less traumatic during inspection, and suitable for detection of large quantities of specimens, it can be used in the clinical diagnosis of malignant tumor patients. It has good application value in monitoring and prognosis evaluation. [0003] The latest statistics from the Chinese Center for Disease Cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/531G01N33/535
Inventor 卢仁泉郭林
Owner 上海永昶医学诊断用品有限公司
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